[A] Difference plot showing reliable discrimination between the wild-type sequence (blue), the c.35 G>C mutation (brown), and the c.37G>T mutation (pink). The Type-it HRM PCR Kit ensured reproducible discrimination with high confidence — without the need for optimization. [B] The c.35 G>C mutation could not be resolved from the wild-type even after extensive optimization of Mg2+ concentration and cycling parameters when using the HRM master mix from Supplier R. The mutation affects the amino acid in position 12, resulting in p.G12A and in position 13, resulting in p.G13C.